Metabolic syndrome, formerly known as syndrome X, is an intermediate state between normal metabolism and type 2 diabetes mellitus. It is an emerging epidemic that has been defined as a series of metabolic risk factors predisposing people to heart and coronary disease and chronic renal failure, among other disease states. Metabolic syndrome is associated with abdominal obesity, atherogenic dyslipidemia, hypertension, proinflammatory and prothrombotic state, with or without glucose impairment, i.e. diabetes. Symptoms are related to high triglyceride levels, low high density lipoprotein levels, increased blood pressure or hypertension and increased glucose levels, and inflammation symptoms. Each of these characteristics is a significant risk factor for development of vascular dysfunction and cardiovascular, renal and hepatic diseases (Eckel R. H et al in Lancet, 2005, 365: 1415) Metabolic syndrome increases the risk of premature death, therefore, effective and affordable strategies to help reduce cardiometabolic alteration factors and to control the syndrome are important goals to benefit a large population being under high risk.
Currently the main approaches to treat people with metabolic syndrome are aimed at reducing the underlying causes, treating hypertension and other cardiovascular risk factors, and control insulin resistance. Treatment of metabolic syndrome includes lifestyle changes, along with drug treatment. In addition to treating metabolic syndrome implementing dietary guidelines and adequate physical activity to address the many underlying mechanisms of metabolic syndrome, a series of medications designed to control the altered levels of metabolites associated with manifestation of the disease state have been developed, incorporating various biologically active nutritional compounds aimed at the same goal. Functional knowledge of nutrient impact, especially amino acids and polyphenols in diet on different molecular targets, makes possible to develop a composition of naturally occurring compounds to treat obesity, metabolic syndrome, diabetes and aging. Some concepts and patents related to the subject of the present invention are described below.
Lodder and Cassis (Lodder R A and Cassis L A in US Patent 2015/9060962 Jun. 23, 2015) patented pharmaceutical compositions comprising D-tagatose with or without stilbene or stilbenoid, or derivatives thereof, to prevent or treat atherosclerosis, metabolic syndrome, obesity or diabetes. Silbenoids such as resveratrol (3, 5, 4-trihydroxy-trans-stilbene) which is a type of natural phenol and a phytoalexin produced in plants was applied during the response to pathogen attack or other injury. Large amounts of resveratrol can be found in different types of berries, grapes, cranberries, etc. (Fremont in Life Sci, 2000, 66: 663). Prior to that patent, Baur et al. (J. A. Baur et al in Nature, 2006, 444 (7117). 337) and Do et al. (Do G M et al in Biochem Biophys Res Commun, 2008, 374 (1). 55) reported the effect of resveratrol to improve health and survival of mice on diets high in calories, control alterations in cholesterol levels, among other capabilities. Another compound chemically related to resveratrol, pterostilbene known as 3,5-dimethoxy-4′hydroxystilbene, is characterized by displaying similar effects to resveratrol.
Sirtuins are enzymes with histone deacetylase and some transcription factor activity that regulate metabolic pathways involved in development of cardiovascular diseases, aging and stress resistance (Jiang W J Biochem. Biophys. Res. Commun., 2008, 373 (3): 341). It is considered that resveratrol directly or indirectly activates SIRT1 (NAD-dependent sirtuin-1 deacetylase) and PGC-1 (gamma receptor coactivator activated by peroxisome proliferator), and affects functioning of mitochondria (Lagouge M. et al in Cells, 2006, 127 (6): 1109; Alcain F J and Villalba J M in Expert Opin Ther Pat, 2009, 19 (4): 403; Beher D. et al in Chem Biol Drug Des, 2009, 74 (6): 619). In cells treated with resveratrol, there is an increase in SOD2 (MnSOD) action (superoxide dismutase 2, mitochondrial, also known as manganese-dependent superoxide dismutase) reducing superoxide, which involves resistance to mitochondrial dysfunction, permeability transition, and death by apoptosis in several diseases. It has been found that resveratrol also acts as a GPER agonist (GPR30) (G protein-coupled estrogen receptor 1, also known as G protein-coupled receptor 30) (Prossnitz E. R. and Barton M. in Mol. Cell. Endocrin. 2014, 389 (1-2): 71). A possible action mechanism of resveratrol can be attributed to autophagy modulation (autophagocytosis—aimed at eliminating dysfunctional mechanism components) (Ghosh H. S. et al in PLoS ONE, 2010, 5 (2). e9199). Laboratory animals have shown the positive effects of resveratrol as an antidiabetic treatment (Baur J A et al in Nature, 2006, 444 (7117): 337; Lagouge M. et al in Cells, 2006, 127 (6). 1109.). This compound acts as PPAR gamma agonist (gamma receptor activated by peroxisome proliferator (PPAR-γ or PPARG), also known as glitazone receptor, or R1C3 (nuclear receptor subfamily 1, group C, member 3) is a type II nuclear receptor), considered as a pharmacological target for treatment of type 2 diabetes (L. Wang et al in Biochem Pharmacol 2014, 92 (1): 73). Peroxisomes are organelles that may cause disease and aging processes while not properly functioning. The ability to stimulate the activity of endothelial nitric oxide synthase (eNOS) and inhibit platelet aggregation is attributed to resveratrol (Vita J. A. Duffy S J in Curr Opinion Lipid, 2003, 14 (1). 21; Olas B. and. Wachowicz B. in Platelets, 2005, 16 (5): 251).
Plant extracts may be considered as antioxidant source, resveratrol analogs. US Patent 2016/9,278,104, Mar. 8, 2016 for Romero T., established the use of Nelumbo nucifera leaf extract to reduce and/or eliminate one or more risk factors associated with metabolic syndrome. The composition considers the presence of creatinine—a nitrogenated organic acid much like amino acid structure located in muscles and nerve cells of different living organism. It is synthesized naturally in the liver, pancreas and kidneys from amino acids such as arginine, glycine and methionine at a rate of one gram of creatine per day. It is considered as an immediate and direct vector to transport ATP and supply energy to muscle myofibrils.
Gokaraju G. R. et al., in US Patent 2016/9,241,964, Jan. 26, 2016 establishes the composition of phytochemical fractions for pharmaceutical use or as a dietary supplement derived from Sphaeranthus indicus and/or Garcinia mangostana. The composition is applied for obesity, metabolic syndrome, diabetes and other metabolic disorder control, prevention and treatment, and also to regulate energy expenditure, prevention of coronary artery and abdominal aorta atherosclerotic plaques, increase insulin sensitivity, control of glucose levels, triglyceride levels and balance glucose levels in mammals. Extracts which contain amino acids among other components are mentioned; however, those are not disclosed.
It is widely known that amino acids are an important part in the synthesis of various proteins involved in human metabolism. This invention is considered to be the reason for which the composition can be selected for the treatment and prevention of metabolic syndrome, obesity, diabetes, and aging.
Varfolomeev S. D. and Gurevich K. G. (in Russian Chemical Bulletin, 2001, 50 (10): 1709) performed biocomputer analysis demonstrating that glycine amino acid is most frequently found (37.5%) in defined positions of primary sequences in different enzyme families. Authors attribute this property to the role of glycine in enzyme architecture: being an amino acid with the chiral atom which allows movement and flexibility to protein chains. This demonstrates the importance of glycine for metabolism. The following 5 amino acids are aspartic acid (12.9%), cysteine (6.7%), histidine (6.2%) and arginine (5.5%). Cysteine is responsible for maintaining the conformation of various proteins due to the formation of sulfur bridges. Histidine and arginine like aspartic acid are common in enzyme active sites by playing a role in the catalytic site as core agents and electrophiles.
In the case of arginine (Wu G. A. B. et al in Curr Opin Clin Nutr Metabol Care, 2000, 3 (1): 59) its ability to help reduce insulin resistance is reported, allowing a decrease in the amount of insulin in diabetes treatments, increase glucose tolerance and improve insulin sensitivity in type 2diabetes mellitus.
Cysteine is the limiting substrate for glutathione synthesis—responsible for protecting cells against viruses, bacteria, fungi, carcinogens, as well as against other disease states. Oral cysteine applied without any other components is not recommended because it is rapidly catabolized to the gastrointestinal tract passing to a toxic state. Cysteine has antioxidant properties, due to the ability of thiols to participate in redox reactions. These cysteine antioxidant properties are mostly expressed in tripeptide glutathione. Availability of oral glutathione is insufficient, thus it must be biosynthesized from the amino acids that constitute it—cysteine, glycine and glutamic acid, from which cysteine is the limiting substrate. Different trials supplemented by laboratory tests have shown that aging is directly associated with a progressive decrease in plasma cysteine and glutathione intracellular concentration. This decrease leads to age-related oxidative stress. Cysteine supplementation above normal diet reduces the various aging processes by helping the bone and muscular systems, reducing inflammation and levels of cytokines (L. Wang et al. In Biochem. Pharmacol. 2014, 92 (1): 73).
Control of cellular proliferation is essential to the proper functioning of any organism. Alteration of this regulation is the cause of diseases such as cancer—with unlimited and uncontrolled cell proliferation due to genetic mutations. Conversely a loss of the ability of cell proliferation is one of the factors causing aging.
Extracellular control of cell division may be performed by cell cycle mitogens, several growth factors as well as by survival factors. Mitogens are proteins that stimulate cell division, counteracting intracellular stop mechanisms (Rb) that block cycle progression.
The growth of any organism or organ depends on both growth and cell division. If cells should be divided without growing, they would become smaller each time. Growth factors stimulate cell growth (cell mass increase) by promoting synthesis and inhibition of proteins and other macromolecules. Cell growth does not depend on cell cycle. For example, nerve and muscle cells grow especially after cell division. Like most mitogens, growth factors bind to cell surface receptors which then activate different intracellular signaling pathways, which may induce: increased protein synthesis or a decreased protein degradation. All this will lead to cell growth.
Survival factors promote cell survival by suppressing intracellular death programs or apoptosis. These signal need from other cells for survival help cells survive only when and where required. Neurons for example, are produced in excess and they compete each other in obtaining growth factors and those with the largest amount will survive. As mitogens and growth factors, survival factors tend to bind to cell membrane receptors which activate intracellular signaling pathways that inhibit cell death, usually by regulating proteins belonging to the Bcl-2 family. Some survival factors are IL-3, SCF, IGF-1, which are mainly secreted by the liver in response to signals from growth hormone (GH).
Based on the above and for the purpose of solving above mentioned problems, the object of present invention is to provide the use of a formulation consisting of: resveratrol, glycine, arginine and cysteine, setting the range of component concentrations based on the study of Artemia salina model. What is patented in this case is the use of the formulation, and the method of its application for treatment of diseases. The formulation has the novel effect to stimulate proliferation of cell cultures and lymphocytes in vitro, without genotoxicity evidence, revealing the ability to decrease formation of visceral fat observed in healthy laboratory animals induced to diabetes, controllability of glycemia index with less insulin or without insuline shown in the model of alloxan-induced diabetes, cytoprotective effect of treatment presented in the pancreas of animals treated with alloxan, correction of symptoms of metabolic syndrome and obesity in patients treated during a 6-month period. In the examples described below it is demonstrated that the formulation has a revitalizing ingredient that act at the level of energy production, restoration of oxygen metabolism and reactivation of various enzymatic cycles related to sugar, fatty acid and cholesterol metabolism, increasing the energy available in the cells for increased functionality and vitality expressed in terms of increased proliferation. Examples demonstrate that when said formulation is applied in animals and humans, altered functions related to negative factors of diabetes, obesity and/or metabolic syndrome are normalized. According to these examples, treatment with the designed formulation is a method to control aging, metabolic syndrome, obesity and diabetes.